Process for the preparation of synthetic dl-tocopherols



Patented Dec. 3, 1946 UNITED STATES PATENT oFl-"lcs PROCESS FOR THE PREPARATION OF SYNTHETIC DL-TOCOPHEROLS Paul Karrer, Zurich, and m Isler, Basel, Switlerland, assignors to Hoflmann-La Roche Inc., Nutley, N. 1., a corporation of New Jersey No Drawing. Application August 12, 1941, Serial No. 406,554. In Switzerland March 31. 1938 Claims.

' our application Serial No. 231,846, filed September 26, 193a.

Evans and collaborators (Mem. Univ. California, vol. 8, year 1927) discovered that a nutritive factor which is indispensable for spermatogenesis in male rats and the successful completion of an existing pregnancy in iemale rats is contained in wheat germ oil and various ioodstufls. They named the new biological i'actor vitamin E and described a biological method oi'determination using female rats. Evans, Emerson and Emerson (J. Biol. Chem., vol. 113, year 1936, page 319, and vol. 122, year 1937, pa e 99), succeeded in isolating three distinct chemical substances from wheat germ oil and various other vegetable oils which are responsible for the vitamin E action oi the starting materials. These closely related compounds were named pand iv-tocopherols.

Investigations by Fernholz (J. Amer. Chem. Soc.,

vol. 59, year 1937, page 1154; vol. 60, year 1938, page 700), Karrer and collaborators (Helvetica Chimica Acta, vol. 20, year 1937; page 1422; vol. 21 year 1938. page 309), Bergel, Todd and collaborators (Biochem. J., vol. 31, year 1937, page 2257; J. Chem. Soc., year 1938, page 253), as well as John and collaborators (Zeitschriit iiir physiologische Chemie, vol. 250, year 1937. P ge 11; vol. 252, year 1938, Pages 201, 208) confirm and supplement the knowledge of the first-named investigators regarding tocopherols.

Natural a-tocopherol was characterized by the empirical formula CzsHsoOs, by an allophanate melting at 158 C., by a p-nitrophenylurethane melting at 131 C., and by a sublimate oi dumhydrouuinone on thermal decomposition. p-Tocopherol possesses the empirical formula Cal-14:02, yields an allophanate melting at 143- 144" C. and a aublimate oi trimethylhydroquinone on thermal decomposition. An allophanate melting at 135 C. was produced from 'Y-tocopherol which has the empirical formula CaHuOz.

'It has now been found that byreacting trimethylhydroquinone or dimethyihydroqulnone with phytol under acid conditions, and Dreierably in the presence of acid condensing agents, com- 5 pounds can be obtained which contain a heterocyclic ring and which must be regarded as chromane derivatives of the general formula CHQCH CH CHCH CHgCH|CHCHsCH CH CHCH 0 Ha Ha Ha Ha wherein two of the R radicals are methyl and the other is methyl or hydrogen. The reaction may, tor example, be carried out in the presence of anhydrous zinc chloride, formic acid or while passing gaseous hydrogen chloride through the reaction mixture. It has also been found particularly suitable to employ a combination of zinc chloride and hydrogen chloride.

The new compounds are the racemates, with respect to carbon atom 2 oi the chromane ring, of tocopherols obtained from natural raw materials. They are light yellow, slightly viscous oils which, in the cold, gradually reduce alcoholic silver nitrate solution. On heating the reduction proceeds quickly. These oils dissolve in concentrated sulphuric acid with a yellow color and the solutions fluoresce intensively alter a iew hours exposure to ultra violet light. The solution or the compounds in chloroform yields a deep dark brown coloration on the addition of tetranitromethane, which gradually clears up. On thermal decomposition the compounds form subiimates oi durohydroquinone or trimethylhydroquinone. When rats so far kept on a vitamin E-iree diet were given these synthetic compounds, it could be established that they rendered it possible for a litter 01' healthy young rats to be born exactly as did tocopherols isolated from natural products.

The new compounds are to be used as pharmaceutical preparations or as starting materials for the manuiacture 0t pharmaceutical preparations.

Example 1 "4 parts by weight of phytol, 2.5 parts by weight of trimethylhydroquinone and 1 part by weight of anhydrous zinc chloride are heated to 180 C. in a current of nitrogen for half an hour. The suspended matter gradually gives a homogeneous brown solution whereby a little hydrogen chloride escapes and a small quantity of trlmethylhydroquinone sublimes. The product is now cooled, precipitated with water, a lot oi ether added, the ether solution washed with water, a solution of caustic soda, and water, dried and the solvent evaporated. The extract is dissolved in petroleurn ether and adsorbed on an aluminium-oxide column. The chromatogram is developed with much petroleum ether. The upper uniformly grey zone contains the condensation product. This is eluted by a mixture of methanol and ether and purified by the preparation or a second chromatogram. The following condensation product is thereby obtained in a pure state:

CH! 03 on. on -C10Hu The new compound from trimethylhydroquinone and phytol contains 80.9 per cent of carbon and 11.8 per cent oi hydrogen. The determination according to Zerewitinofi indicates the presence of one active hydrogen atom, i. e., a phenolic hydroxyl group. The compound possesses a characteristic absorption spectrum with a maximum absorption at 294 pp. and a minimum absorption at 256 a l. It forms various crystalline derivatives, such as a p-nitrophenyl-urethane melting at 131 C., an allophanate melting at 172 C., and a 3,5-dinitro benzoic ester melting at 63 C. On administering a dose of 3 mg. to a rat kept on a vitamin E-iree diet, the full action of the pure vitamin was observed.

Example 2 4 parts by weight of phytol, 2.2 parts by weight of trimethylhydroquinone and 1 part by weight of anhydrous zinc chloride are suspended in 10 parts by weight of decalin and then heated to 150-160 C. for one hour while stirring and introducing carbon dioxide. Th trimethylhydroquinone dissolves completely, whereby the liquid becomes yellowish brown. The product is cooled, water and ether added with stirring, the ether solution washed successively with water, a solution or caustic soda, hydrochloric acid, and water, dried with sodium sulphate and the ether evaporated. The residue is adsorbed on an aluminiumoxide column. After development of the chromatogram with a good deal of petroleum ether, the column is uniformly grey. At the foot thereof there is a violet zone. The grey main zone is eluted with a mixture of methyl alcohol and ether (8:1), the solvent evaporated and the extract iurther purified by the preparation of a second chrcmatogram. The product is then distilled in a molecular distillation apparatus and the compound described in the previous example obtained. This compound was given in doses 0! 3 and 10 mg. each to four rats which had been kept on a diet free from vitamin E. All the animals carried the pregnancy to term and brought forth healthy young.

Example 3 11 parts by weight of phytol, parts by weight of trimethylhydroquinone and 60 parts by weight of anhydrous formic acid are boiled together for four to flve hours under reflux. The product is then diluted with water, extracted with ether. the ether evaporated and the residue saponified with an alcoholic solution of sodium ethylate or iii] alcoholic potash. When the Example 4 4 parts by weight oi phytol, 2.5 parts by weight of 3,5-dimethylhydroquinone and 1 part by weight of anhydrous zinc chloride are heated to 180 C. for one-half an-hour while stirring and introducing carbon dioxide. The suspended matter gradually fuses gi i a o eneous brown melt. After cooling, the product is treated with ether and water, the ether solution washed with potash and water, and the ether residue adsorbed on an aluminium oxide column. The chromatogram is developed with much petroleum ether. The whole of the lower part of the column is whitish-grey. The top layer of aluminium oxide, which is hardly colored, is removed and the whitish-grey principal zone eluted with a mixture of methyl alcohol and ether (3:1). The solvent is evaporated in vacuo and the residue distilled in a molecular distillation apparatus. The compound is a yellow, slightly viscous oil, nn' =1.501. A sublimate oi trimethylhydroquinone results on thermal decomposition. An allophanate melting at 143 C, is obtained with cyanic acid in benzene solution. A 5 mg. dose of this compound wa biologically fully active.

Example 5 4 parts by weight of phytol, 2.5 parts by weight of 3,5-dimethylhydroqulnone and 1 part by weight of zinc chloride are suspended in 10 parts by weight of decaline and then heated to C. for one hour while heating and introducing carbon dioxide. The m-xylohydroquinone dissolves completely. The fluid becomes yellowish-brown. The product is cooled, water and ether added while stirring, the ether solution washed with water, potash, hydrochloric acid and water, dried with sodium sulphate and the ether evaporated. The residue is adsorbed on an aluminium oxide column. After developing the chromatogram with much petroleum ether, the column appears almost uniformly yellowish-grey. At the bottom there is a small yellow zone. The yellowish-grey principal layer is eluted, the solvent evaporated and the residue purified by the preparation or a second chromatogram. The product is then distilled in a molecular distillation apparatus and the compound described in Example 4 is obtained.

Example 6 4 parts by weight of phytol, 2 parts by weight of 2,5-dimethyl hydroquinone and 1 part by weight of zinc chloride are heated to C. for one-half hour while stirring and introducing carbon dioxide. When the material has become homogeneous, it is cooled, treated with ether and water and the ether solution washed with potash, then with water, the ether layer separated, dried, and the ether removed by distillation. The residue is taken up in small volume of low boiling petroleum ether and chromatogramed on an aluminium oxide column. By eluting the yellowish top zone with a mixture of ether and methanol,

an oil is obtained which reduces a neutral solution of an alcoholic solution of-silver nitrate on boiling. The material corresponds to the structure:

Its aliophanate melts at 154 C.

Example 7 11 parts by weight of phytol. 5 parts by weight of 2,3-dimethyl hydroq'uinone; 60 parts by weight of anhydrous formic acid are refluxed together for four to five hours. The product i then diluted in water, extracted with ether, the ether evaporated, the residue saponifled with an alcoholic solution 01' sodium methylate or alcoholic potash. when the alcohol has been distilled off, the residual product is extracted with ether and the ether solution washed and dried. The ether is removed and the residue is taken up in a small quantity of low boiling petroleum ether and chromatogramed on an aluminium oxide column. This gives a yellow-brownish layer in the upper part of the absorption column which on elution contains the expected condensation product consisting of a viscous oil with strong reducing action. The analysis corresponds to the formula CacHaOr. It possesses an active hydrogen atom and corresponds to the structure:

on c

H: The allophanate thereof melts at 146 C. when tested on rats which had been kept on a vitamin E-i'ree diet, the compound was fully active in a dose oi mg.

Example 8 2 parts by weight oi zinc chloride, 10 parts by weight of phytol, and 5 parts by weight of trimethyl hydroquinone are dissolved in a mixture of parts by volume of ether and 20 parts by volume of benzol, maintaining a temperature of 45-55? C. Dry hydrochloric gas is passed through the mixture until complete saturation which requires from two to four hours. The reaction liquid is then washed with water, then with hydrochloric acid, again with water, then with sodium CHE:

-wlmicmdnoucmcnmcncnl o om em al n,

wherein two of the R radicals represent a. methyl group and the third R radical a radical selected from the group consisting of hydrogen and methyl radicals, by condensing a methyl substituted hydroquinone of the formula wherein two of the radicals R represent a methyl group and the third 8 radical represents a radical selected from the group consisting of hydrogen and methyl adicals, by condensing a methyl substituted hydroquinone of the formula wherein two of the R radicals represent a methyl group and the third R radical represents a radical selected from the group consisting 0! hydrogen and methyl radicals, with phytoi in the presence of zinc chloride, and recovering a tocopherol product from the reaction mixture thus obtained.

3. In a process for the manufacture of a tocopheroi product, the steps of producing a tocopherol of the formula 7 Ho OH:

R I (CH1)1CH(OHI)ICH(6HI)ICHCHJ o n. on, H. H:

wherein two of then radicals represent a methyl group and the third R radical represents a radical selected from the group consisting of hydrogen and methyl radicals, by condensing a methyl substituted hydroquinone of the formula wherein two of the it radicals represent a methyl group and the third R. radical representsa radical selected from the group consisting of hydrogen andmethylradicais. with phytol inthepreaence of nine chloride. a solvent. while passins gaseous hydrosen chloride through the mixture, and recoverins a tocopherol product from the reaction mixture thus obtained.

4. In a process for the copherol product, the steps oi copherol of the formula producing an e-to- -(CHs)sCH(OHl)sUB(GHQICHOHI Ks Ha Ha HI Ha which comprises condensing trimethyihydroquinone with phytol' under acid conditions, and

manuiacture o! a tov recovering a tocopherol product from the reaction mixture thus obtained.

5. In a process for the manufacture of a tocopherol product, the steps of producing an e-tocopherol oi the formula which comprises condensing trimethylhydroquinone with phytol in the presence of zinc chloride, and recovering a tocopherol product from the reaction mixture thus obtained.

e. In a process for the manufacture or a tocopherol product, the steps oi producing a. to-

copherol oi' the formula r no on. OH -(om).cmcmnomcnmoncn- 0 HI H! H: H:

which comprises condensing 3,5'-dimethyihydroquinone with phytol under acid conditions. and recovering. a tocopheral product from the reaction mixture thus obtained.

'I. In a process for the manufacture of a to- 8 oopherol product. the steps 0! producinl a tocopherol of the iormula OH! no em 0 b-(on|).cn(emncn(cnmcncm I 0 ts. bin in m which comprises condensing SJ-dimethrlhrdroquinone with phytol in the presence 0! zinc chloride, and recovering a tocopherol product from the reaction mixture thus obtained.

8. In a process for the manufacture of a tocophercl product, the steps oiproducing a tocopherol o! the formula CHI He bonmomomhcn(camcncm 0 Us Kl as B! which comprises condensing 2,5-dimethylhydroquinone with phytol under acid conditions, and

recovering a tocopherol product from the reaction mixture thus obtained.

- 9. In a process for the manuiacture of a tocopheroi product.the steps of producing a tocopherol oi the formula HO Kl hammer: onmomonmcnon.

0 on. m n. in

Certificate or Correction PAUL KARRER ET AL.

It is hereby certifiai that error appears in the patent issued correction as follows:

2412968 read 2411968; and t should be read with -thiscorrection therein that the same may the case in thePatent Oifice.

25th day of February, A. D. 1947.

inventors on December 3, 1946, requiring right handoorner, for the patent number Letters Patent to the record of Signed and sealed this to the above named In the ant, upper at the said conform LESLIE FRAZER,

ence of nine chloride. a solvent. while passins gaseous hydrosen chloride through the mixture, and recoverins a tocopherol product from the reaction mixture thus obtained.

4. In a process for the copherol product, the steps oi copherol of the formula producing an e-to- -(CHs)sCH(OHl)sUB(GHQICHOHI Ks Ha Ha HI Ha which comprises condensing trimethyihydroquinone with phytol' under acid conditions, and

manuiacture o! a tov recovering a tocopherol product from the reaction mixture thus obtained.

5. In a process for the manufacture of a tocopherol product, the steps of producing an e-tocopherol oi the formula which comprises condensing trimethylhydroquinone with phytol in the presence of zinc chloride, and recovering a tocopherol product from the reaction mixture thus obtained.

e. In a process for the manufacture or a tocopherol product, the steps oi producing a. to-

copherol oi' the formula r no on. OH -(om).cmcmnomcnmoncn- 0 HI H! H: H:

which comprises condensing 3,5'-dimethyihydroquinone with phytol under acid conditions. and recovering. a tocopheral product from the reaction mixture thus obtained.

'I. In a process for the manufacture of a to- 8 oopherol product. the steps 0! producinl a tocopherol of the iormula OH! no em 0 b-(on|).cn(emncn(cnmcncm I 0 ts. bin in m which comprises condensing SJ-dimethrlhrdroquinone with phytol in the presence 0! zinc chloride, and recovering a tocopherol product from the reaction mixture thus obtained.

8. In a process for the manufacture of a tocophercl product, the steps oiproducing a tocopherol o! the formula CHI He bonmomomhcn(camcncm 0 Us Kl as B! which comprises condensing 2,5-dimethylhydroquinone with phytol under acid conditions, and

recovering a tocopherol product from the reaction mixture thus obtained.

- 9. In a process for the manuiacture of a tocopheroi product.the steps of producing a tocopherol oi the formula HO Kl hammer: onmomonmcnon.

0 on. m n. in

Certificate or Correction PAUL KARRER ET AL.

It is hereby certifiai that error appears in the patent issued correction as follows:

2412968 read 2411968; and t should be read with -thiscorrection therein that the same may the case in thePatent Oifice.

25th day of February, A. D. 1947.

inventors on December 3, 1946, requiring right handoorner, for the patent number Letters Patent to the record of Signed and sealed this to the above named In the ant, upper at the said conform LESLIE FRAZER, 

